Protective effects and mechanism of Panax Notoginseng saponins on oxidative stress-induced damage and apoptosis of rabbit bone marrow stromal cells.

OBJECTIVE

To investigate the effects and possible mechanism of Panax Notoginseng saponins (PNS) on oxidative stress-induced damage and apoptosis in bone marrow stromal cells (BMSCs).

METHODS

BMSCs were isolated and cultured from 2-month-old New Zealand rabbits by the density gradient centrifugation combined with adherent method. The third passage cells were used for subsequent experiments. Oxidative stress was induced in cultured BMSCs by H(2)O(2) (0.1 mmol/L). BMSCs were pretreated with 25-200 μg/mL PNS for 4 h before H(2)O(2) treatment. Proliferation of BMSCs was observed using MTT assay. Alkaline phosphatase (ALP) activity, as an index of early osteoblastic differentiation, was determined with an ALP assay kit. Flow cytometry was used to observe the apoptosis of BMSCs by staining with annexinV-FITC/propidium iodide. Oxidative stress level was examined by reactive oxygen species (ROS) assay. The protein expressions of Bax, Bcl-2 and Caspase-3 in BMSCs were analyzed by Western blotting.

RESULTS

PNS had different concentration-dependent effects on proliferation and osteoblast differentiation of BMSCs induced by H(2)O(2). A PNS concentration of 100 μg/mL was determined as the optimal effective concentration. PNS markedly attenuated H(2)O(2)-induced apoptosis rate from 41.91% to 14.67% (P<0.01). PNS significantly decreased ROS level induced by H(2)O(2) (P<0.01). Furthermore, pretreatment with PNS significantly reversed H(2)O(2)-induced inhibition of Bcl-2 expression and augmentation of Bax and Caspase-3 expression (P<0.01).

CONCLUSIONS

PNS had a protective effect on oxidative stress-induced damage and apoptosis in cultured rabbit BMSCs through scavenging ROS and regulating the Bcl-2/Bax pathway.